Piezo1 Antibody - #DF12083

Figure 5. Mechano-stress initiates the positive loop of NF-kB and periostin via PIEZO1 (A) Immunoblotting results of three control and three degenerated human NP tissues. C, control; D, degenerated. (B) Quantitation of Ca2+ influx changes in hNPCs after 5 mM Yoda1 stimulation (left panel), and representative fluorescence images at indicated time points (right panel). Yoda1 was administrated at 10 s. (C) Representative images of Fluo-8 AM in hNPCs cultured on different hydrogels. Scale bar = 100 mm. (D) Representative b-Gal staining images of hNPCs treated with vehicle or Yoda1 for 24 h (left panel), and quantitation of b-Gal+ senescent cells (right panel). n = 3 biologically independent samples. Scale bar, 100 mm. (E–G) qPCR results of senescence (E), SASP (F), and matrix metabolism (G) markers in hNPCs treated with vehicle or Yoda1 for 24 h. n = 3 biologically independent samples. (H) Representative immunostaining images of NF-kB p65 (red) in hNPCs under indicated treatments for 30 min. DAPI (blue) indicates nuclei. (I) Quantification of p65 localization in (H), presented as mean ± SD. In total, 237 cells from the vehicle group and 232 cells from the Yoda1 group were analyzed. *Represents the statistical analysis was conducted in terms of the proportion of N > C between the two groups and the p < 0.05. n = 3 biologically independent samples. Scale bar = 25 mm. (J) Immunoblotting results of hNPCs treated with vehicle or Yoda1 for 48 h. (K and L) Representative b-Gal staining images of hNPCs under indicated conditions for 24 h (K), and quantitation of b-Gal+ senescent cells (L). n = 3 biologically independent samples. Scale bar, 100 mm. (M) Relative mRNA expression of the matrix metabolism and senescence markers and POSTN in hNPCs under indicated conditions. n = 3 biologically independent samples. (N and O) Representative T2WI image (N) and MRI Pfirrmann grade (O) of rat tail IVDs with indicated administration. n = 9 discs.

Fig. 6 Characterization results of integrin β1 and piezo 1 expressions in H9c2 cells on the PA gels with stiffness of 23.9 and 60.1 kPa. A Fluorescence images and B statistical histogram of integrin β1 in the H9c2 cells on the PA gels with different stiffness (n>3). C fluorescence images and D statistical histogram of piezo 1 in H9c2 cells on the PA gels with different stiffness (n>3). ns, no significant difference, *p 

Figure 6. UVR-induced dynamic alterations of lymphatic structure and function regulated by YAP1/Piezo1 and collagen. (A) Image of Masson staining. Magnification of microscopic image: 100 ×. (B) Image of VE-CAD (VE-CAD with brown tint), (C) Piezo1 (Piezo1 with brown color), and (D) YAP1 (YAP1 with brown color) immunochemistry staining in the dermis. Statistics of the area of VE-CAD (E), YAP1 (F), and Piezo1 (G) per area in immunochemistry staining. The expression of YAP1 after eight weeks of UVR was the least. When normal tissue progressed to AK, it exceeded normal skin. When it moved into cSCC, the positive area increased rapidly. Piezo1 represented the opposite trend compared with YAP1. (H) The trend lines of YAP1, LVs, and Piezo1 value. (I)Statistics of collagen thickness. (J) Statistics of the grade of collagen disorganization. The amount of collagen decreased. The collagen bundle thickened and disorderly arranged in a mass accumulation. Magnification of microscopic image: 100 ×. (K) The potential lymphatic-centered immune microenvironment markers in different skin stages induced by UVR.*, p < 0.05 and **, p

Figure 2 Validation and phenotype of Piezo1 IntL-CKO mice fed with high-fat diet. (A) Body weight of 14- to 16-week-old male Piezo1loxp/loxp and Piezo1 IntL-CKO mice fed with HFD for 10 weeks (n=6/group). (B) IPGTT and associated area under the curve (AUC) values of 14- to 16-week-old male Piezo1loxp/loxp and Piezo1 IntL-CKO mice fed with HFD (n=6/group). (C) Gcg mRNA levels in the ileal mucosa of 14- to 16-week-old male Piezo1loxp/loxp and Piezo1 IntL-CKO mice fed with HFD (n=6/group). (D) The plasma GLP-1 level in 14- to 16-week-old male Piezo1loxp/loxp and Piezo1 IntL-CKO mice fed with HFD (n=6/group). (E) Double immunofluorescent staining of Piezo1, and GLP-1 in the ilea of 14- to 16-week-old male Piezo1loxp/loxp and Piezo1 IntL-CKO mice fed with HFD (n=6/group). (F) Representative western blots are shown for indicated antibodies in the ileal mucosa (n=6/group). (G) Body weight after 7 consecutive days infusion of saline or Ex-4 (100 µg/kg body weight) in 14- to 16-week-old male Piezo1loxp/loxp and Piezo1 IntL-CKO mice fed with HFD (n=6/group). (H, I) IPGTT (H) and ITT (I) and associated area under the curve (AUC) values after consecutive infusion of saline or Ex-4. (J) Gcg mRNA levels in the ileal mucosa (n=6/group) after consecutive infusion of saline or Ex-4. (K) The plasma GLP-1 level after consecutive infusion of saline or Ex-4 (n=6/group). Data are represented as mean ± SEM. Significance was determined by Student’s t test for comparison between two groups, and by one-way ANOVA for comparison among three groups or more,

Figure 1. Piezo1 was up-regulated in the articular cartilage of OA patients and rats. (a,c) Representative images of HE, Safranin-O/Fast Green staining of intact and damaged areas in OA patients’ articular, Sham, and OA rat cartilage, and OARSI scores (n = 5). (b,d) Immunohistochemistry staining of intact and damaged areas in OA patients’ articular, Sham, and OA rat cartilage, and the percentage of Piezo1 positive cells (n = 5). (e) Immunofluorescence staining and quantified results of Piezo1 in chondrocytes exposed to mechanical strain for 0 h, 6 h, 12 h, 24 h, and 48 h (n = 3). (f,g) RT-qPCR and Western blot analysis of Piezo1 in chondrocytes exposed to mechanical strain for 0 h, 6 h, 12 h, 24 h, and 48 h. * p < 0.05; ** p < 0.01; ns, no significant differences. Mann–Whitney U test for (a,c), Student’s t-test for (b,d), one-way ANOVA with Bonferroni’s test for (e–g); scale bar: 100 μm.

Figure 1. Piezo1 was up-regulated in the articular cartilage of OA patients and rats. (a,c) Representative images of HE, Safranin-O/Fast Green staining of intact and damaged areas in OA patients’ articular, Sham, and OA rat cartilage, and OARSI scores (n = 5). (b,d) Immunohistochemistry staining of intact and damaged areas in OA patients’ articular, Sham, and OA rat cartilage, and the percentage of Piezo1 positive cells (n = 5). (e) Immunofluorescence staining and quantified results of Piezo1 in chondrocytes exposed to mechanical strain for 0 h, 6 h, 12 h, 24 h, and 48 h (n = 3). (f,g) RT-qPCR and Western blot analysis of Piezo1 in chondrocytes exposed to mechanical strain for 0 h, 6 h, 12 h, 24 h, and 48 h. * p < 0.05; ** p < 0.01; ns, no significant differences. Mann–Whitney U test for (a,c), Student’s t-test for (b,d), one-way ANOVA with Bonferroni’s test for (e–g); scale bar: 100 μm.